Abstract
T cells are an immune cell lineage that play a central role in protection against pathogen infection. Antigen, in the form of pathogen-derived peptides, stimulates the T-cell receptor (TCR), leading to activation of the transcription factor, nuclear factor kappa B (NF-κB). The subsequent NF-κB-dependent gene expression program drives expansion and effector differentiation of antigen-specific T cells, leading to the adaptive anti-pathogen immune response. The cell surface TCR transmits activating signals to cytosolic NF-κB by a complex signaling cascade, in which the adapter protein Bcl10 plays a key role. We have previously demonstrated that TCR engagement leads to the formation of cytosolic Bcl10 clusters, called POLKADOTS, that provide a platform for the assembly of the terminal signaling complex that ultimately mediates NF-κB activation. In this chapter, we describe the methods utilized to visualize the formation of TCR-induced POLKADOTS and to study the temporal association between POLKADOTS formation and nuclear translocation of the NF-κB subunit, RelA/p65.
Key words
- T cells
- TCR
- Bcl10
- POLKADOTS
- NF-κB
- RelA/p65
- Confocal microscopy
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Acknowledgments
Supported by grants from the US National Institutes of Health (Al057481 to B.C.S.), Center for Neuroscience and Regenerative Medicine (CNRM) (to B.C.S.), and predoctoral fellowships (to S.P.) from the American Heart Association (10PRE3150039) and the Henry M. Jackson Foundation. The views expressed are those of the authors and do not necessarily reflect those of the Uniformed Services University or the Department of Defense.
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Paul, S., Schaefer, B.C. (2015). Visualizing TCR-Induced POLKADOTS Formation and NF-κB Activation in the D10 T-Cell Clone and Mouse Primary Effector T Cells. In: May, M. (eds) NF-kappa B. Methods in Molecular Biology, vol 1280. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2422-6_12
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DOI: https://doi.org/10.1007/978-1-4939-2422-6_12
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